Mahatma gandhi university

Structural organization of DNA and RNA: Organization of pro and eukaryotic genomes, nucleotypes (diversity in genome size), coding regions, non coding regions, introns and repetitive sequences; DNA and RNA polymerases, restriction enzymes, gene cloning and in situ hybridization of DNA .DNA replication: Mutation, DNA repair; Site directed mutagenesis; Polymerase chain reaction and its applications .Gene expression: Gene expression in prokaryotes and Ekaryotes, Transcription and Translation; Reverse transcription; Regulation of gene expression; Transposable elements; Expression of foreign DNA in pro and Eukaryotes (35 hrs)


References:

• 
  Benchamin Lewin. 2003. Genes. (8^th ed.). Prentice Hall Inc.
  

• 
  Brown, T.A. (Ed.). 2002. Essential Molecular Biology. Vol.2. Oxford University Press, New York.
  

• 
  Harvey Lodish, Arnold Berk, Paul Matsudaira, Chris A. Kaiser, Monty Krieger,
  Matthew P. Scot, Lawrence Zipursky and James Darnell. 2004. Molecular Cell Biology. (5^th ed.). Freeman and Co., New York
  

• 
  Jacquelin.C. Black. 2004. Microbiology: Principles and Explorations. (5^th ed.) John Wiley and Sons Inc.
  

• 
  James Watson, Mark Zoller, Michael Gilman and Jan Witkowski. 1993. Recombinant DNA (2^nd ed.). Scientific Academic Press, Ney York
  

• 
  James Watson, Tania Baker, Stephen Bell, Alexander Gann, Michael Levine, and Richard Losick. 2004. Molecular Biology of the Gene. (5^th ed.) Prentice Hall Inc.
  

• 
  Jones, P.G. and Sutton, J.M. 1977. Plant Molecular Biology Essential Techniques. John Wiley & Sons, New York
  

• 
  Michael T. Madigan and John M. Marktinko. 2006. Brock Biology of Microorganisms (11^th ed). Prentice Hall Inc.
  

• 
  Pelczar, M.J. Jr., Chan, E.C.S., Krieg, N.R. 1993. Microbiology. (5^th ed.). Tata McGrew Hill Pub. Co. Ltd., New Delhi.
  

• 
  Pommerville. 2004. Alcamo’s Fundamentals of Microbiology (7^th ed). Jones and Bartlett Publishers Inc.
  

• 
  Tortora Christine, Case, L. and Berdell R. Funke. 2003. Microbiology: An Introduction (8th ed.). Benjamin Cummings Publishers Inc.
  

PHYTOM2M002PP2- Laboratory course-II

PRACTICALS (based on Phytom 05 and Phytom 06) (150 hrs)

1. 
   Plant physiology: Determination of leaf extract (cell sap) pH; Estimation of relative water content in leaves; Extraction of pigments; Separation and identification of pigments (chlorophylls, carotenoids and xanthophylls) by chromatography; Growth analysis: Net primary productivity (NPP), Relative growth rate (RGR), Net assimilation rate (NAR) and Leaf weight ratio (LWR).
   

2. 
   Estimation of chlorophyll: Spectrophotometric assay of total chlorophyll, chlorophyll-a and chlorophyll-b.
   

3. 
   Biochemistry: Estimation of total sugar, reducing sugars, lipids, phenolic compounds, terpenes, terpenoids, amino acids and proteins; Assay of enzymes: protease, amylase, invertase and succinic dehydrogenase; Electrophoresis: Protein
   

4. 
   Microbiology: Staining of bacteria: simple, Gram’s staining, spore staining, etc.; Media preparation: Nutrient agar, blood agar, etc.; Cultivation of microbes, bacterial motility and colony characteristics; Biochemical tests for bacterial identification; Study of anti-bacterial effects of plant extracts; Sterility tests for Ayurvedic concepts.
   

5. 
   Ecology: Community analysis for quantitative characteristics through quadrat method: quadrat sampling, minimum size and number of quadrat, measurement of density, abundance, frequency, dominance and importance value index (IVI).
   

PHYTOM3M003PC7 Cell Biology and Biotechnology (100 hrs)

1. 
   Ultra structure of plant cell: Organization and functions of cell; Cell membrane and cell organelles: cytosol, membranes, cytoskeleton, endoplasmic reticulum, nucleus, nucleolus, chromosomes, golgi complex, lysosomes, peroxisomes, mitochondria and chloroplast; Nucleo-cytoplasmic transport (10 hrs)
   

2. 
   Cell cycles: Phases of cell cycle, check point of cell cycle, extra cellular cell signals (hormones, cytokinines and growth factors), signal receptors, photoreception; Chromosomes and cell division: chromatin structure (histones, nucleosomes and fibers) and chromosome replication, mitosis (nuclear envelope, centromere, telomeres) meiosis (chromosome pairing, synapsis and movement, meiotic spindle), cytokinesis (20 hrs)
   

3. 
   Chromosomal (numerical and structural) variation: Euploids, aneuploids and polyploids; β-chromosomes; structural aberrations; Experimental induction of numerical and structural changes in chromosomes, role of chromosomal variation in evolution and plant breeding (15 h)
   

4. 
   Recombinant DNA technology: DNA sequence analysis: gene libraries, gene cloning, cutting and joining of DNA; Transformation, vectors, laboratory synthesis of genes, nucleic acid hybridization; Plant improvement: improvement of plants through manipulation of phenotypic traits; protoplast isolation and fusion; genetic engineering and transgenic plants. Economic, Social and ethical implications: Intellectual and material property rights; Technology transfer; International trade; Ethical and social concerns regarding the use of genetically engineered organisms or their products in agriculture and medicine (20 hrs)
   

5. 
   Applications of biotechnology: Plant tissue culture: Totipotency of cells; concept and general composition of culture media; cell, tissue and organ culture; Somatic embryos, somaclonal variation; control of differentiation, growth and flowering; Haploid production, embryo rescue; Micropropagation Applications in: Pharmaceuticals (production of therapeutic proteins, drugs and nutritional supplements), Medicine (molecular diagnosis of disease, monoclonal antibodies, gene therapy, biotransformation, vaccine production), Forensic science (blood typing and DNA finger printing), Industry (enzyme engineering, biocatalysts, biopolymers, biosensors), Environment management (bioremediation), Genetic conservation (storage of DNA, cells, pollen, tissues and organs by in vitro methods), and Food (enhanced keeping quality, nutritional superiority, fermentation products (35 hrs)
   

References:

• 
  Agutter, P.S. 1991. Between nucleus and cytoplasm. Chapman and Hall, London
  

• 
  Balasubramanian, D., Bryce, C.F.A., Dharmalingam, K., Green J. and Jayaraman, K. (eds) 1996. Concepts in Biotechnology. Universities Press (India) Ltd., Hyderabad
  

• 
  Callow, J.A., Ford-Lloyd, B.V. and Newbury, H.J. 1997. Biotechnology and Plant GeneticResources Conservation and Use. CAB International Oxon
  

• 
  Chirikjian, J.G. (ed). 1995. Biotechnology: Theory and Techniques (Vol. 1&2). Jones & Barlett Pub., Boston
  

• 
  Clark, M.S. and Wall, W.J. 1996. Chromosomes the Complex Code. Chapman and Hall, London
  

• 
  Dodds, J.H. and Roberts, L.W. 1995. Experiments in Plant Tissue Culture (3^rd ed). Cambridge University Press, Cambridge
  

• 
  Driel, R.V. and Otte, A.P. (eds). 1997. Nuclear Organization, Chromatin Structure and Gene Expression. Oxford University Press, Oxford
  

• 
  Elgin, S.C.R. (ed) 1995. Chromatin Structure and Gene Expression. IRL Press, Oxford
  

• 
  Endow, S.A. and Glover, D.M. (eds). 1998. Dynamics of Cell Division. Oxford University Press, Oxford
  

• 
  Falconer, D.S. and Mackay, T.F.C. 1996. Quantitative Genetics. (4^th edn). Longman, Essex
  

• 
  Gamborg, O.L. and Phillips, G.C. (eds). 1995. Plant Cell, Tissue and Organ culture. Narosa Pub. House, New Delhi
  

• 
  Hancock, J.T. 1997. Cell signaling. Longman Essex
  

• 
  John, B. 1990. Meiosis. Cambridge University Press, Cambridge
  

• 
  George, E.F. and Sherrington, P.D. 1984. Plant Propagation by Tissue culture. Exegetics Limited, Hants
  

• 
  Kingsman, S.M. and Kingsman, A.J. 1998. Genetic Engineering. Blackwell Scientific Pub., London
  

• 
  Kumar, U. 1999. Methods in plant tissue culture. Agro Botanica, Bikaner
  

• 
  Kyte, L. and Kleyn, J. 1996. Plants from Test tubes: An Introduction to Micro-propagation (3^rd ed.). Timber Press, Portland
  

• 
  Murray, R.M. (ed). 1991. Advanced Methods in Plant Breeding and Biotechnology. CAB International Oxon
  

• 
  Ravishankar, G.A. and Venkatesh, L.V. 1997. Recent Advances in Biotechnological Applications of Plant Tissue and Cell culture. Oxford & IBH Pub. Co. Pvt. Ltd., New Delhi
  

• 
  Stebbins, G.L. 1971. Chromosomal evolution in higher plants. Edward Arnold, London
  

• 
  Tewari, K.K. and Singhal, G.S. 1997. Plant Molecular Biology and Biotechnology. Narosa Pub. House, New Delhi
  

• 
  Walker, J.M. and Gingold, E.B. 1993. Molecular Biology and Biotechnology (3^rd ed.). Royal Society of Chemistry, Cambridge
  

PHYTOM3M003PC8 Phytochemistry (100 hrs)

1. 
   Phytochemical techniques: Cold and hot extraction methods, liquid-liquid extraction techniques, liquid-carbon dioxide extraction, concentration and evaporation techniques, lyophilisation; TLC, preparative TLC, PC, column chromatography, gel-chromatography, affinity chromatography, ion-exchange chromatography, gas-liquid chromatography, high performance liquid chromatography, high performance thin layer chromatography and colorimetric analysis of extracts; finger printing of extracts and estimation of bioactive molecules (20hrs)
   

2. 
   Bioactive secondary metabolites: Steroids: Occurrence and distribution in plants, saponins, sapogenins and steroids; Isolation, structure elucidation and synthesis of bioactive steroids such as cholesterol, diosgenin, estrone, estrodiol, etc.; Terpenoids: Occurrence and distribution in plants, essential oils, aroma chemicals, mono and sesquiterpenoids, their use in flavour and perfumery industry, diterpenes, triterpenes, isolation and characterization of terpenes, their synthesis; Alkaloids: occurrence and distribution in plants, bioactive alkaloids-isolation and structure elucidation of alkaloids such as atropine, quinine, papaverine, thebaine, vincristine, etc.; Anthocyanidin: occurrence and distribution in plants, isolation and characterization of anthocyanins, chalcones, flavones, isoflavones, chromones, coumarins; structure elucidation of quercetin, kaempferol, etc. (25hrs)
   

3. 
   Spectroscopy: UV/Visible, IR, 1H and 13 C NMR, 2D NMR and MS; application of spectroscopic techniques in structural elucidation of secondary metabolites (20 hrs)
   

4. 
   Instrumentation: Analytical Methods: Light and election microscopy, tissue printing cytochemical localization, immunological methods (production of monoclonal and polyclonal antibodes, agglutination and precipitation tests, immuno diffusion assays, immuno electrophoresis, radio immunological assay), radioactive isotopes and autoradiography, photometry and radiometry, atomic spectroscopy, GC-MS, LC-MS, centrifugation, electrophoresis, IEF and 2D electrophoresis, joint selective electrodes, enzymes, medicinal analysis (25 hrs)
   

5. 
   Applications: Applications of biomolecular chemistry in plant systematics, plant physiology, medicine and pharmaceuticals, forensic science, environmental science, biotechnology, herbal and modern drug industries, food flavour and cosmetic industries (10 hrs)
   

References:

• 
  Agarwal, P.K., Thakur, R.S. and Bansal, C.M. 1989. Carbon-13 NMR of Flavonoids. Elsevier Science Publishers, Amsterdam
  

• 
  Anderson. J.W. and Beardall. J. 1991. Molecular Activities of Plant Cells. Blackwell Scientific Publications, Oxford
  

• 
  Apps, D.K. and Tipton, K.F. 1995. Essays in Biochemistry, Portland Press, London
  

• 
  Athkins, P. and Jones, I. 1999. Chemical Principles the Quest for Insight. W.H Freeman and Co. New York.
  

• 
  Atlas, R.M.1995. Principles of Microbiology. Mossy, St.Lois
  

• 
  Atta-ur-Rahman. 1989. One and Two Dimensional NMR Spectroscopy (1^st ed.). Elsevier, New York
  

• 
  Braithwaite, A. and Smith, F.J. 1996. Chromatographic Methods (5^th ed) Blackie Academic & Professional London
  

• 
  Budzikiewicz, H., Djerassi, C. and Williams, D.H. 1968. Mass Sectrometry of Organic Compounds. Holden-Day, San Francisco, CA
  

• 
  Dennis, D.T and Turpin, D.H. (Eds.). 1990. Plant physiology, Biochemistry and Molecular biology, Longman Scientific and Technical Essex
  

• 
  Derome, A.E. 1987. Modern NMR Techniques for Chemistry Research (1^st ed.). Pergamon Press, New York
  

• 
  Dey, P.M. and Harborne, J.B. (Eds.). 1997. Plant Biochemisty, Academic Press, New York
  

• 
  Harborne, J.B. 1984. Phytochemical Methods (2^nd ed). Chapman and Hall, London.
  

• 
  Howarth, J. 1998. Core Organic Chemistry, John Wiley & Sons Chichester
  

• 
  Kemp, W. 1993. Organic Spectroscopy (3^rd ed.). ELBS, Hong Kong
  

• 
  Lea, P.J. and Leegood, R.C. 1993. Plant Biochemistry and Molecular Biology. John Wiley & Sons, West Susses
  

• 
  Mann J, Davidson, R.S Hobbs J.B Banthorpe.D.V and Harborne J.B. 1994. Natural Products. Longman Scientific and Technical Essex
  

• 
  Reed, R. Holmes, J., Weyers, J. and Jones, A.1998. Practical Skills in Biomolecular Sciences. Longman, Essex
  

• 
  Reid, P.D. and Pont-Lezica, R.F. (Eds.). 1992. Tissue Printing. Academic Press, New York
  

• 
  Schwedt, G. 1997. The Essential Guide to Analytical Chemistry. John Wiley & Sons, New York
  

• 
  Silverstein, R.M., Bassler, G.C. and Morrill, T.C. 1981. Spectroscopic Identification of Organic Compounds (4^th ed.). John Wiley, New York
  

• 
  Smith, P.M. 1976. The Chemotaxonomy of Plants. Edward Arnold, London
  

• 
  Solomon, P.H. and Nakanishsi, K. 1998. Infrared Absorption Spectroscopy (2^nd ed.). Adams E (Pr) Inc.
  

• 
  Thomspn, R.H. (Ed.) .1985. The Chemistry of Natural Products. Blackie London
  

• 
  Turner, P.C. Mel.ennan, A.G. Bates, A.D. and White, M.R. 1997. Instant Notes in Molecular Biology. Bios Scientific Publishers, Oxford.
  

• 
  Wilson, K. and Walker, J. (Eds). 1994. Principles and Techniques of Practical Biochemistry (4^th ed) Cambridge University Press, Cambridge.
  

• 
  Whitehead, D. and Bowers, W.S. 1983. Natural Products for Innovative Pest Management. Pergamon, Oxford
  

PHYTOM3M003PC9 Pharmacognosy (100 hrs)

1. 
   Pharmacognosy: Definition, scope and applications in herbal medicine Classification and identification of drugs: Need for classification of drugs; classical (taxonomic, morphological, organoleptic, therapeutic); microscopy and modern (biogenetic) criteria for classification of powdered drug, methods for documentation of raw drugs (35 hrs)
   

2. 
   Processing of plant drugs: Methods of collection, process and storage of medicinal and aromatic plants; purification of raw drugs; factors causing drug contamination, methods of storage of drugs (20 hrs)
   

3. 
   Drug administration in Indian systems of medicine: The holistic concept of drug administration; description of Sapta padarthas in Dravya guna; (20 hrs)
   

4. 
   Single plant drugs and formulations in Ayurveda, Siddha and Unani; classical and modern means of drug administration (20 hrs)
   

5. 
   Pharmacognosy of crude drugs: based on market survey including adulterants and substitutes (5 hrs)
   

References:

• 
  Anonymous. 1978. The Ayurvedic Formulary of India. Govt. of India, New Delhi
  

• 
  Anonymous. 1989. Formulary of Siddha Medicine. The Indian Medical Practitioners’ Co-operative Pharmacy and Stores Ltd., Chennai
  

• 
  Anonymous. 1999. The Ayurvedic Pharmacopoeia of India. Vol. I (1 & 2). Ministry of Health and Family Welfare, Govt. India, New Delhi.
  

• 
  Chauhan, M.G. and Pillai, A.P.G. 2005. Microscopic Profile of Powdered Drugs Used in Indian Systems of Medicine. Institute of Ayurvedic Medicinal Plant Sciences, Jamnagar.
  

• 
  Daljithsimha, K. 1974. Unani Dravyaguna Darshana. Ayurvedic and Tibbi Academy, Lucknow
  

• 
  ICMR. 2003. Quality Standards of Indian Medicinal Plants (Vol.1). Indian Council of Medical Research, New Delhi
  

• 
  Jackson, B.P. and Snowdon, D.W. 1992. Atlas of Microscopy of Medicinal Plants, Culinary herbs and Spices. CBS Pub., New Delhi
  

• 
  Kameswara Rao, C. 2000. Database of Medicinal Plants. KSCST, Bangalore
  

• 
  Narayana Aiyer, K. and Kolammal. M. 1963. Pharmacognosy of Ayurvedic Drugs (12 vol.). University of Kerala, Thiruvananthapuram
  

• 
  Said, H.M. (Ed.). 1969. Hamdard Pharmacopoeia of Eastern Medicine. Institute of Health and Tibbi Research, Karaachi
  

• 
  Sivarajan, V.V. and Indira, Balachandran. 1994. Ayurvedic Drugs and their Plant Sources. Oxford & IBH Pub. Co. Pvt. Ltd. New Delhi
  

• 
  Trease, G.E. and Evans, W.C. 1983. Pharmacognosy (12^th ed.). Bailliere Tindall, London
  

• 
  Vaidya, B. 1982. Some Controversial Drugs in Indian Medicine. Chaukambika Orientalia, Varanasi
  

• 
  Wallis, T.E. 1997. Text Book of Pharmacognosy (5^th ed) CBS Publishers & Distributors, Delhi
  

PHYTOM3M003PP3 Laboratory course-III

PRACTICALS (based on Phytom-07 to Phytom-9) (150 hrs)
Phytochemistry: Estimation of fixed oil from fruit kernel (e.g. Percentage yield of coconut oil in copra); Determination of saponification value, Iodine value and acid value of fixed oil; Estimation of nitrogen in plant parts using Kjeldahl’s method; Estimation of the alkaloid piperine from Pepper, vasicine from Adhatoda leaves and carbohydrates in plant samples using Duboi’s method; Thin Layer Chromatography of the Petroleum ether extract of Cinnamon bark/Clove/Nutmeg kernel/Turmeric/Cardomom seeds; Paper Chromatography of 70% aqueous alcoholic extract of Hibiscus leaves/Curry leaves/ Muringa leaves for amino acids (spray reagent: Ninhydrin); Isolation of Starch from Potato; Isolation and estimation of caffeine from tea leaves; Paper Chromatography of monosaccharides from fruits such as Mango/ Jack fruit /Orange; Estimation of ascorbic acid from gooseberry.
Pharmacognosy: Study of physical evaluation of crude drugs: determination of optical rotation and refractive index of drug samples (Castor oil); determination of Rf-value by Thin Layer Chromatography; determination of total ash value, acid soluble ash value and water soluble ash value of crude drug samples; morphological/taxonomic description (including morphological description of useful plant parts) and identification of important medicinal and aromatic plants of Kerala; histochemical/chemical tests for identifying various plant components such as suberin, cutin, mucilage, pectin, starch, protein, oils, alkaloids, tannin, calcium oxalate and calcium carbonate, in various crude drug samples; organoleptic evaluation of 25 crude drug samples: colour, odour, taste and other features; tests for detection of organic acids in: fruits of tamarind and lemon, leaves of Oxalis corniculata, etc.; Anatomical studies: killing and fixing of tissues, tissue dehydration, paraffin infiltration, mounting, microtomy, staining and mounting; anatomical study of selected plants: Cinnamomum verum, Mimusops elengi, Azadirachta indica and Saraca asoca; Micrometry: determination of cell size and area using micrometers and cameralucida; determination of stomatal index and frequency.

Raw drug resources: Collection and submission of raw drug samples (25 samples)


PHYTOM4M004PE1 Pharmacology, Clinical trial and Biostatistics (100 hrs)

1. 
   Pharmacology and Ethnopharmacology: Definition, scope and applications in herbal medicines; Ethnopharmacology: Pharmacological evaluation of drugs from ethnomedicine (Strychnos nux-vomica, Rauvolfia serpentina and Digitalis species); Importance of ethnopharmacological studies. Criteria for pharmacological evaluation of drugs: Absorption, distribution, elimination, pharmacokinetics and drug reception; Drug abuse and iatrogenic diseases .Pharmacological activity of plant drugs: Examples of plant drugs with effects on the autonomous and central nervous systems, cardiovascular system and the gastrointestinal system; Plant drugs with antimicrobial activity; Plant chemicals in modern pharmacology: Biochemistry and pharmacology of atropine, caffeine, ephedrine, opioids, taxol, Vinca alkaloids; synthetic substitutes for therapeutically active plant constituents; drug improvement by structure modification and biotransformation .